Real-time monitoring of cellular superoxide anion release in THP-1 cells using a catalytically amplified superoxide dismutase–based microbiosensor
Reactive oxygen species (ROS) including the superoxide anion (O 2 •− ) are typically studied in cell cultures using fluorescent dyes, which provide only discrete single-point measurements. These methods lack the capabilities for assessing O 2 •− kinetics and release in a quantitative manner over lon...
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Veröffentlicht in: | Analytical and bioanalytical chemistry 2024-09, Vol.416 (21), p.4727-4737 |
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Sprache: | eng |
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Zusammenfassung: | Reactive oxygen species (ROS) including the superoxide anion (O
2
•−
) are typically studied in cell cultures using fluorescent dyes, which provide only discrete single-point measurements. These methods lack the capabilities for assessing O
2
•−
kinetics and release in a quantitative manner over long monitoring times. Herein, we present the fabrication and application of an electrochemical biosensor that enables real-time continuous monitoring of O
2
•−
release in cell cultures for extended periods (> 8 h) using an O
2
•−
specific microelectrode. To achieve the sensitivity and selectivity requirements for cellular sensing, we developed a biohybrid system consisting of superoxide dismutase (SOD) and Ti
3
C
2
T
x
MXenes, deposited on a gold microwire electrode (AuME) as O
2
•−
specific materials with catalytic amplification through the synergistic action of the enzyme and the biomimetic MXenes-based structure. The biosensor demonstrated a sensitivity of 18.35 nA/μM with a linear range from 147 to 930 nM in a cell culture medium. To demonstrate its robustness and practicality, we applied the biosensor to monitor O
2
•−
levels in human leukemia monocytic THP-1 cells upon stimulation with lipopolysaccharide (LPS). Using this strategy, we successfully monitored LPS-induced O
2
•−
in THP-1 cells, as well as the quenching effect induced by the ROS scavenger N-acetyl-
l
-cysteine (NAC). The biosensor is generally useful for exploring the role of oxidative stress and longitudinally monitoring O
2
•−
release in cell cultures, enabling studies of biochemical processes and associated oxidative stress mechanisms in cellular and other biological environments.
Graphical Abstract |
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ISSN: | 1618-2642 1618-2650 1618-2650 |
DOI: | 10.1007/s00216-024-05437-z |